Large-scale production of CMP-NeuAc and sialylated oligosaccharides through bacterial coupling (original) (raw)

Abstract

A large-scale production system of cytidine 5′-monophospho-_N_-acetylneuraminic acid (CMP-NeuAc) and sialyloligosaccharides was established by a whole-cell reaction through the combination of recombinant Escherichia coli strains and Corynebacterium ammoniagenes. For the production of CMP-NeuAc, two recombinant E. coli strains were generated that overexpressed the genes of CMP-NeuAc synthetase and CTP synthetase, respectively. C. ammoniagenes contributed to the formation of UTP from orotic acid. CMP-NeuAc was accumulated at 27 mM (17 g/l) after a 27-h reaction starting with orotic acid and _N_-acetylneuraminic acid. When E. coli cells that overexpressed the α-(2 → 3)-sialyltransferase gene of Neisseria gonorrhoeae were put into the CMP-NeuAc production system, 3′-sialyllactose was accumulated at 52 mM (33 g/l) after an 11-h reaction starting with orotic acid, _N_-acetylneuraminic acid, and lactose. Almost no oligosaccharide byproducts other than 3′-sialyllactose were observed after the reaction. The production of 3′-sialyllactose at a 5-l jar fermenter scale was almost the same as that at a beaker scale, which indicated the high potential of the 3′-sialyllactose production on an industrial scale.

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Authors and Affiliations

1. Tokyo Research Laboratories, Kyowa Hakko Kogyo Co., Ltd., 3-6-6, Asahimachi, Machida, Tokyo 194-8533, Japan e-mail: endo.tetsuo@kyowa.co.jp Tel.: +81-42-7252555 Fax: +81-42-7268330, , , , , , JP
   T. Endo, S. Koizumi, K. Tabata & A. Ozaki

Authors

1. T. Endo
2. S. Koizumi
3. K. Tabata
4. A. Ozaki

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Received: 9 July 1999 / Received revision: 17 September 1999 / Accepted: 10 October 1999

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Endo, T., Koizumi, S., Tabata, K. et al. Large-scale production of CMP-NeuAc and sialylated oligosaccharides through bacterial coupling.Appl Microbiol Biotechnol 53, 257–261 (2000). https://doi.org/10.1007/s002530050017

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• Issue date: March 2000
• DOI: https://doi.org/10.1007/s002530050017

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